Klasyczne leczenie modyfikujące oraz leczenie biologiczne a ryzyko rozwoju nowotworów w reumatologii

  W leczeniu chorób reumatycznych jednym z problemów jest ryzyko rozwoju nowotworów, zarówno w kontekście wysokiej aktywności zapalnej, jak i w odniesieniu do nowoczesnych terapii. Współcześnie coraz szersze stosowanie leków modyfikujących przebieg choroby, a także nowych leków biologicznych, nasuwa pytanie o bezpieczeństwo takiej terapii. Ta publikacja jest próbą odpowiedzi na pytanie, czy w świetle publikowanych danych terapie biologiczne mają związek z większym ryzykiem rozwoju nowotworów. Znacząca większość dostępnych danych informuje o dużym profilu bezpieczeństwa stosowania leków biologicznych. Wykazują one, że ryzyko rozwoju nowotworów nie różni się istotnie od ryzyka populacyjnego. Forum Reumatol. 2016, tom 2, nr 1, 32–38 Słowa kluczowe: leki biologiczne; nowotwory; ryzyko nowotworu

Hypermethylation of CpG islands and shores around specific microRNAs and mirtrons is associated with the phenotype and presence of bladder cancer

PURPOSE To analyze the role and translational potential for hypermethylation of CpG islands and shores in the regulation of small RNAs within urothelial cell carcinoma (UCC). To examine microRNAs (miR) and mirtrons, a new class of RNA located within gene introns and processed in a Drosha-independent manner. EXPERIMENTAL DESIGN The methylation status of 865 small RNAs was evaluated in normal and malignant cell lines by using 5-azacytidine and microarrays. Bisulfite sequencing was used for CpG regions around selected RNAs. Prognostic and diagnostic associations for epigenetically regulated RNAs were examined by using material from 359 patients, including 216 tumors and 121 urinary samples (68 cases and 53 controls). Functional analyses examined the effect of silencing susceptible RNAs in normal urothelial cells. RESULTS Exonic/UTR-located miRs and mirtons are most susceptible to epigenetic regulation. We identified 4 mirtrons and 16 miRs with CpG hypermethylation across 35 regions in normal and malignant urothelium. For several miRs, hypermethylation was more frequent and dense in CpG shores than islands (e.g., miRs-9/149/210/212/328/503/1224/1227/1229), and was associated with tumor grade, stage, and prognosis (e.g., miR-1224 multivariate analysis OR = 2.5; 95% CI, 1.3-5.0; P = 0.006). The urinary expression of epigenetically silenced RNAs (miRs-152/328/1224) was associated with the presence of UCC (concordance index, 0.86; 95% CI, 0.80-0.93; ANOVA P < 0.016). CONCLUSIONS Hypermethylation of mirtrons and miRs is common in UCC. Mirtrons appear particularly susceptible to epigenetic regulation. Aberrant hypermethylation of small RNAs is associated with the presence and behavior of UCC, suggesting potential roles as diagnostic and prognostic biomarkers

Integrated Epigenome Profiling of Repressive Histone Modifications, DNA Methylation and Gene Expression in Normal and Malignant Urothelial Cells

Epigenetic regulation of gene expression is commonly altered in human cancer. We have observed alterations of DNA methylation and microRNA expression that reflect the biology of bladder cancer. This common disease arises by distinct pathways with low and high-grade differentiation. We hypothesized that epigenetic gene regulation reflects an interaction between histone and DNA modifications, and differences between normal and malignant urothelial cells represent carcinogenic events within bladder cancer. To test this we profiled two repressive histone modifications (H3K9m3 and H3K27m3) using ChIP-Seq, cytosine methylation using MeDIP and mRNA expression in normal and malignant urothelial cell lines. In genes with low expression we identified H3K27m3 and DNA methylation each in 20–30% of genes and both marks in 5% of genes. H3K9m3 was detected in 5–10% of genes but was not associated with overall expression. DNA methylation was more closely related to gene expression in malignant than normal cells. H3K27m3 was the epigenetic mark most specifically correlated to gene silencing. Our data suggest that urothelial carcinogenesis is accompanied by a loss of control of both DNA methylation and H3k27 methylation. From our observations we identified a panel of genes with cancer specific-epigenetic mediated aberrant expression including those with reported carcinogenic functions and members potentially mediating a positive epigenetic feedback loop. Pathway enrichment analysis revealed genes marked by H3K9m3 were involved with cell homeostasis, those marked by H3K27m3 mediated pro-carcinogenic processes and those marked with cytosine methylation were mixed in function. In 150 normal and malignant urothelial samples, our gene panel correctly estimated expression in 65% of its members. Hierarchical clustering revealed that this gene panel stratified samples according to the presence and phenotype of bladder cancer

Global epigenetic profiling in malignant and normal urothelial cell lines

Epigenetic regulation of gene expression IS commonly altered in human cancer. Numerous alterations of DNA methylation and microRNA expression have been described in bladder cancer and these reflect the biology and phenotype of the disease. This common disease arises by distinct pathways with low and high grade differentiation. Epigenetic gene regulation is stabilized within a chromatin state by maintainers such as histone modification or DNA methylation. I hypothesized that epigenetic gene regulation reflects an interaction between histone and DNA modifications, and differences between normal and malignant urothelial cells represent carcinogenic events within bladder cancer. To test this hypothesis I profiled two histone modifications thought to be repressive (H3K9-3M and H3K27-3M) using ChIP-Seq, cytosine methylation using MeDIP and epG Isl and microarrays and mRNA expression in normal urothelial cells and cell lines that represent non invasive and invasive tumours. In genes with low expression I identified associated H3K27-3M and DNA methylation each in 20-30% of genes and both marks in 5% of genes. H3K9-3M was detected in 5-10% of genes but was not associated with overall expression. DNA methylation was more closely related to gene expression in malignant than normal cells. H3K27-3M was the epigenetic mark most specifically con-elated to gene silencing. I identified a panel of genes with cancer speci fie-epigenetic mediated aberrant expression including those with reported carcinogenic functions and members potentially mediating a positive epigenetic feedback loop. Pathway enrichment analysis revealed genes marked by H3K9-3M were involved with cell homeostasis, those marked by H3K27-3M mediated pro-carcinogenic processes and those marked with cytosine methylation were mixed in function. Hierarchical clustering revealed that gene panel stratified samples according to the presence and phenotype of bladder cancer. finally, the last part of my thesis concentrates on identifying epigenetic events associated with chemoresistance in bladder cancer. Currently, acquired resistance to drugs is the major obstacle in the successful treatment of advanced UCC. In addition, many tumours initially respond to chemotherapy, but subsequently develop secondary resistance. In order to identify epigenetic mechanisms associated with chemoresistance, I developed eisplatin resistant cell lines with 2 different approaches and investigated genome wide dysregulation of DNA methylation and microRNA expression in these. Resistant cells generated with different methodology and consequently with different stress applied, had very few common characteristics. They were found to have distinct profiles of microRNAs expression with more differentially expressed microRNAs (11=39) in the resistant cells derived from heterogeneous cell populations when compared to those from single cell clones (n=16). Only 8 microRNAs were shared between the resistant cells (including 5 with increased and 3 with decreased expression). Established resistant cells were also characterised by very distinct methylation profiles. The heterogeneous population of resistant cells were found to have the highest number of enriched and decreased in 5MC probes when compared to the cell lines generated from the single clones. Only 0.1 % of 5MC enriched and 0.02% 5MC decreased probes were shared between the established resistant cells. In summary, this Thesis illustrates that DNA methylation, histone modifications and microRNA expression are important regulatory mechanisms altered within UCc. These represent potential biomarkers and therapeutic targets that could be used to overcome treatment resistance owing to the reversible nature of their modifications.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Global epigenetic profiling in malignant and normal urothelial cell lines

Epigenetic regulation of gene expression IS commonly altered in human cancer. Numerous alterations of DNA methylation and microRNA expression have been described in bladder cancer and these reflect the biology and phenotype of the disease. This common disease arises by distinct pathways with low and high grade differentiation. Epigenetic gene regulation is stabilized within a chromatin state by maintainers such as histone modification or DNA methylation. I hypothesized that epigenetic gene regulation reflects an interaction between histone and DNA modifications, and differences between normal and malignant urothelial cells represent carcinogenic events within bladder cancer. To test this hypothesis I profiled two histone modifications thought to be repressive (H3K9-3M and H3K27-3M) using ChIP-Seq, cytosine methylation using MeDIP and epG Isl and microarrays and mRNA expression in normal urothelial cells and cell lines that represent non invasive and invasive tumours. In genes with low expression I identified associated H3K27-3M and DNA methylation each in 20-30% of genes and both marks in 5% of genes. H3K9-3M was detected in 5-10% of genes but was not associated with overall expression. DNA methylation was more closely related to gene expression in malignant than normal cells. H3K27-3M was the epigenetic mark most specifically con-elated to gene silencing. I identified a panel of genes with cancer speci fie-epigenetic mediated aberrant expression including those with reported carcinogenic functions and members potentially mediating a positive epigenetic feedback loop. Pathway enrichment analysis revealed genes marked by H3K9-3M were involved with cell homeostasis, those marked by H3K27-3M mediated pro-carcinogenic processes and those marked with cytosine methylation were mixed in function. Hierarchical clustering revealed that gene panel stratified samples according to the presence and phenotype of bladder cancer. finally, the last part of my thesis concentrates on identifying epigenetic events associated with chemoresistance in bladder cancer. Currently, acquired resistance to drugs is the major obstacle in the successful treatment of advanced UCC. In addition, many tumours initially respond to chemotherapy, but subsequently develop secondary resistance. In order to identify epigenetic mechanisms associated with chemoresistance, I developed eisplatin resistant cell lines with 2 different approaches and investigated genome wide dysregulation of DNA methylation and microRNA expression in these. Resistant cells generated with different methodology and consequently with different stress applied, had very few common characteristics. They were found to have distinct profiles of microRNAs expression with more differentially expressed microRNAs (11=39) in the resistant cells derived from heterogeneous cell populations when compared to those from single cell clones (n=16). Only 8 microRNAs were shared between the resistant cells (including 5 with increased and 3 with decreased expression). Established resistant cells were also characterised by very distinct methylation profiles. The heterogeneous population of resistant cells were found to have the highest number of enriched and decreased in 5MC probes when compared to the cell lines generated from the single clones. Only 0.1 % of 5MC enriched and 0.02% 5MC decreased probes were shared between the established resistant cells. In summary, this Thesis illustrates that DNA methylation, histone modifications and microRNA expression are important regulatory mechanisms altered within UCc. These represent potential biomarkers and therapeutic targets that could be used to overcome treatment resistance owing to the reversible nature of their modifications.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Case report Adult-onset Still’s disease as a mask of Hodgkin lymphoma

Adult-onset Still’s disease is a rare disorder, which creates difficulties in making a proper diagnosis. Ambiguous symptoms and results of auxiliary tests, lack of unequivocal diagnostic tests and the need to exclude other causes of the disease are major problems in clinical practice. A case of a 22-year-old woman with dominated recurrent fever, significantly elevated inflammation markers and arthritis is presented. Based on clinical signs after exclusion of infection, hematological and other reasons, the patient was diagnosed with adult-onset Still’s disease. Standard treatment, with high doses of glucocorticoids and a disease-modifying drug, was applied, without the anticipated effects. The diagnostic tests were conducted again due to the lack of clinical improvement, increase of inflammatory markers and unusual response to treatment. A new symptom of significance, i.e. mediastinal lymphadenopathy, was found. After the histopathological examination of lymph nodes, Hodgkin’s disease was diagnosed and targeted therapy for hematological malignancy was applied

Anchor Effect in Polymerization Kinetics: Case of Monofunctionalized POSS

The effect of the anchoring group on the detailed polymerization kinetics was investigated using monomethacryloxy-heptaisobutyl POSS (1M-POSS). This compound was copolymerized with lauryl methacrylate (LM) as the base monomer, at various molar ratios. The process was initiated photochemically. The polymerization kinetics were followed by photo-DSC and photorheology while the polymers were characterized by nuclear magnetic resonance (NMR), gel permeation chromatography (GPC), transmission electron microscopy (TEM), and differential scanning calorimetry (DSC). For comparison, a methacrylate containing the branched siloxy-silane group (TSM) was also studied. It was found that the modifiers with a bulky substituent have a dual effect on the termination process: (i) At low concentrations, they increase the molecular mobility by increasing the free volume fraction, which leads to an acceleration of the termination and slows the polymerization; while (ii) at higher concentrations, they retard molecular motions due to the &ldquo;anchor effect&rdquo; that suppresses the termination, leading to acceleration of the polymerization. The anchor effect can also be considered from a different point of view: The possibility of anchoring a monomer with a long substituent (LM) around the POSS cage, which can further enhance propagation. These conclusions were derived based on kinetic results, determination of polymerization rate coefficients, and copolymer analysis

Nowy funkcjonalny napełniacz hybrydowy TiO2-SiO2/wielościenny oligomeryczny silseskwioksan, jako potencjalny modyfikator polietylenu

The paper presents preliminary application studies of the innovative TiO2 -SiO2 / POSS filler as a potential low-density polyethylene modifier. It was found that the addition of the filler increases the stiffness and hardness of the composite and slightly changes the intensity of its color.Przedstawiono wstępne badania aplikacyjne innowacyjnego napełniacza TiO2 -SiO2 /POSS jako potencjalnego modyfikatora polietylenu małej gęstości. stwierdzono, że dodatek napełniacza powoduje wzrost sztywności i twardości kompozytu oraz nieznacznie zmienia intensywność jego barwy

Enrichment for DNA bound to H3K9m3 and H3K27m3 around Transcriptions Start Sites (TSS).

<p>Although deep sequencing reads were mapped throughout the genome, there was enrichment around that varied between H3K9m3 and H3K27m3. The average number of reads within the 2 kb window surrounding 33,183 transcription start sites (TSS) is shown.</p